Department of Biochemical Engineering

Determination of the best possible design in CSTR and Batch reactors for the
production of galacturonic acid of pectin present in green banana residues

L.Montilla, S. Mosquera, I, Ordoñez, K, Peña

Icesi Cali University, Colombia

Abstract:
This article seeks galacturonic acid from Musa paradisiaca shell residues. Batch and
CSTR reactors were evaluated for this purpose in order to achieve the best possible
design for this bioprocess. Galacturonic acid was obtained from the pectin present in Musa
paradisiaca shells by enzymatic hydrolysis. The enzyme used for the process was
pectinas 831L (P831L) for which its kinetic parameters were determined by Matlab.
Volumes for batch and CSTR of 0.8607 m 3and 1,7473 m 3 respectivelywere3 obtained.
Finally, based on certain criteria, the batch reactor was chosen as the most suitable for
bioprocessing.

Keywords: Pectina, galacturonic acid, Reactor batch, CSTR reactor, Kinetic parameters

1. Introduction
In Colombia oneof the main problems is the high generation of agro-industrial waste
such as banana peels, where most of the time, they are burned or thrown into
garbage dumps, rivers and ravebs without any prior treatment, however the efficient
use of these wastes for the manufacture of by-products generate income at the
industrial level; since it is one of the agricultural products that are most cultivated in
the Colombian fields; according to statistics from the Ministry of Agriculture and
Rural Development, a study carried out in the period January to September 2010
exported 78,874 tonnes, due to the ignorance of many banana-producing
communities to which knowledge of the usefulness that this waste has about 95% of
the waste generated from bananas is not efficiently exploited , since production
focuses on marketing or as a food option for the home [1]. However, it is possible,,
in recent years it has been studied that they can be a great source for the extraction of
pectin.

Pectin is a specific group of carbohydrate polymers composed largely of D-

galacturonic acid chains that may be bound by bonds (1-4) with side chains of L-
arabinosa and D-galactose, and whose carboxylic groups may be partially
methoxylated or fully neutralized by bases, there is a wide range of esterification
degrees depending on species, tissue and maturity [2].] In general, tissue pectins
have a range of esterification grades ranging from 60 to 90%. Pectin is used as a
gelling agent, thickener, emulsifier and stabilizer in various food products, where it
has been found toyield higher yields,20.68% m/m [3] having a new opportunity in the
banana industry, therefore it is necessary to evaluate what is the best design for the
extraction of pectin that fulfills the purpose of the study by making a comparison
between a Batch or CSTR type reactor.

2.Materials and methods

The company Platanitos Cope provided the wasteable banana peels which are ( 66250
kg /year) of which 20.6% percentage was used for pectin extraction.

2.1 Removing pectin

First the banana peels were washed successively with distilled water to remove traces of
pulp and impurities present on the surface, then the shells were filtered into a liencillo cloth
to remove excess water, followed by this the shells were cut, ground and enzymatic
inactivation was performed for 15 minutes at 95-98oC, then a wash was performed to
remove solid residues, was filtered again to remove excess water and impurities and was led
to a drying at a temperature of 60o C. Then acid hydrolysis was used with HCL at pH of 2.0
and immediately heated to 85oC the mixture along with a mechanical agitation constantly so
that no precipitate will be generated for 60 minutes and immediately heated to 85oC the
mixture along with a mechanical agitation constantly so that no precipitate will be generated
for 60 minutes , immediately filtered to separate the liquid phase with 50% v/v ethanol and
the acidic solution was precipitated to obtain only pectin; finally, it dried at a temperature of
40oC for 2 h.

2.2 Determination of kinetic parameters

2.2.1 Experimental

The enzymatic kinetics of the p830L was estimated thanks to the evaluation of 7 assays with
different substrate concentrations, keeping theenzyme c eleventh constant, the
absorbedancia read in each test was measured, and the different initial rates were
determined, these experimental provided data were used for modeling,and subsequent
determination of cynical parameters. [5]

2.2.2 Mathematical model

The enzyme kinetics for the enzyme p830L was modeled using the generic equation for
inhibition por substrate presented in (Ec 1).

(Ec 1)

2.3.1. D- Galacturonic acid yield

Galacturonic acid performance was taken from the Pectin-to-Galacturonic Acid: Waste-to-
Bioproductsreporttheory.[ 5] using 92% galacturonic acid recovery.

2.3.2. Purification of galacturonic acid:

Purification begins with an ultrafiltration to separate other large molecules

(monosaccharides and oligosaccharides) from hydrolyzing. He then underwent an
electrodialysis separation process with bipolar membranes to separate galacturonic acid by
taking advantage of its negative charge. Subsequently, crystallization was made to obtain
galacturonic acid from the acid solution, resulting from electrodialysis. It was eventually
vacuum filtered to remove methanol and water that was added during the process and dried
in the oven to remove excess water and obtain crystals of pure galacturonic acid. This
procedure is best illustrated in Fig 1.

2.4. Evaluation of bioreactors.

The evaluation of CSTR and Batch reactors was carried out through the approach of the
design equations, under some assumptions taken into account for each type of reactor and
subsequently MATLAB was used to model the design of each reactor, obtaining its volume
and quantity of production per year. So they were compared and decided on the best design
for this particular process. In addition to this, other factors such as the economic and the
advantages and disadvantages of each reactor will betaken into account in order to be able
to choose the bestone. The amount of pectin per year 13250 kg/year and the conversion
(0.92) were left fixed for both reactors..

2.4.1 Reactor Batch

For the design of this reactor, the theoretical equation was used to model an ideal batch
reactor (Ec 2), also took into account the downtime that is designated for preparation,
emptying, filling, cleaning and sterilization.

(EC 2)

2.4.2 Reactor CSTR

For the design of this reactor, the generic equation for substrate inhibition enzymatic kinetics
(EcEc 3)wasused. The equation was also used for an ideal CSTR reactor (Ec 4)..

(Ec 3)
 (Ec 4)

3. Results and discussion

3.1 Determination of cinematic parameters

The pectinasa 831 L enzyme was used in the determination of the experimentaldata, which
works optimally in a pH range of 2.5- 5.5 and a temperature range of 40-55oC.
Based on experimental data obtained in the laboratory and provided for the study, the
enzymatic kinetics data presented in Table 1 were reached. [5]

Table 1. Experimental data on enzymatic kinetics

.
For the determination of the parameters, the results obtained in Table 1 were modeled.
MATLAB mathematical software, as shown in Fig 1. Obtaining the values below:

Vmax= 21,097 uM/min

Km-4.33 g/L
Ks=7.670 g/L
 Fig 1. Experimental data obtained from the initial speed vs the concentration of
substrate used to obtain kinetic parameters.

3.2. Galacturonic acid performance.

According to the information provided the recovery performance of galacturonic acid after
enzymatic hydrolysis was 92%, i.e. the conversion of pectin into galacturonic acid was 92%
[5] which is considered a very good yield since the pectin is not only made up of galactronic
acid, it also contains some neutral sugars such as D-galactose, L-arabinosa and L-
ramanosa [4]. On the other hand, from the purification of the recovered hydrolysation, pure
crystallized galacturonic acid can be obtained, with a purity yield of 98% [6] indicating that
bipolar electrodialysis with membrane and subsequently crystallization turn out to be good
techniques to obtain pure galacturonic acid and ready for commercialization.

3.3 Evaluation of bioreactors

In the evaluation of these two types of bioreactors was used the production of banana peel
residues that was provided by the company Platanitos Cope, 66250 kg/year, of these was
taken only 20% corresponding to the total of the shell without moisture, after a drying
process. The total percentage of pectin in shell, equivalent to 20.6% [3], using cáscara,
2729.5 kg/year, as a total pectin quantity was also taken into account.

3.3.1 Reactor Batch

To obtain the reaction time of the batch reactor was made use of the EC equation [2], it was
evaluated with a conversion of 92% galacturonic acid, yielding for the reactor volume a total
of 0.8607 m3..

3.3.2 Reactor CSTR

For CSTR reactor modeling Ec 7 was used considering the steady state, ec 1substrate
inhibition cinéti ca was also taken into account. It took 25 days as maintenance time, taking
into account emptying, sterilization, drying and subsequent filling; was evaluated for a
conversion of 92% galacturonic acid and atotal volume of 1,7473 m 3was obtained for this
reactor..

Discussion

For the processing of 2729.5 kg/year of extracted pectin, a volume of 0.8067m 3 and an
initial substrate concentration of 10 g/L, fig 3, in a batch reaction time of 25.62 hours, fig 4,
were obtained for the Batch ream processing. As for the CSTR reactor, aminimum required
volumen of 1.7473m3 with an initial concentration of 10 g/L, fig 2, was determined in a
continuous reaction for 340 days, taking into account the maintenance time taken, 25 days.
To carry out the comparison between the two reactors for both the batch reactor and the
CSTR reactor, it was modeled for a 92% conversion and the same amount of processed
pectin.

Fig 2. CSTR reactor volume vs initial substrate concentration

Fig 3. Batch reactor volume vs initial substrate concentration

Fig 4. Batch reactor reaction time vs initial substrate concentration

3.4 Criteria for bioreactor selection

For the selection of the bioreactor, the conditions under which each bioreactor works, as well
as the results obtained after modeling each bioreactor in. Matlab.

For the CSTR bioreactor it was assumed that it has both input and output flows of the
chemostat,so it is considered an open system. In addition, it has a good mix and, both the
temperature and the concentration are identical anywhere in the bioreactor, just like in the
output flow. [7] Finally, taking into account the inhibition by sustrato, in which Ec 1 was
used to determine its kinetics, modeling with ec Ec 7 and the remaining assumptions
named above (Item 2.4),resulted in a volume of1,7473 m3 for 92% withgalacturonic acid
version,

Conversely, the BATCH batch bioreactor has no input or output flows so it is a closed
system. It also does not operate in a steady state, such as CSTR, as properties such as
concentration or temperature may vary over time. On the other hand, this type of bioreactor
has a good mixing, which means that it remains homogeneous. With the help of ec 2
modeling in MATLAB and taking into account the remaining assumptions named above
(Item 2.4) and theinhibition by substrate that it presents, the reaction time (25.62 hours) was
obtained for the same percentage (92%) conversion of galacturonic acid. As well as the
volume (0.8607 m3) of reaction.
Table 2. Advantages and desven

Based on the results obtained and the pros and cons of each bioreactor, it was decided to
work with the BA bioreactor

4. Conclusions

5. References

[1] Banana residues can be efficiently exploited. 04/03/2011, from CARACOL RADIO
https://caracol.com.co/radio/2011/03/04/ecologia/1299231060_434552.html

[2] CALABRANO CALABRANO SARAVIA, RODRIGO DUARTE NASS, EDUARDO VERA

SUAZO. (2014). "PECTIN: CHEMISTRY, SOURCES, EXTRACTION PROCESS,
GELIFICATION AND ITS USE IN THE ELABORATION OF JAMS OR JELLIES AND
OTHER APPLICATIONS. "

[3] A. Vasquez, L.Ruesga,R D'addosio,G Paez and M. Marin. (21-06-2006). Extraction

of pectin from the banana shell (MusaAAB, banana subgroup) clone fed up .
Re.Fac.Agron , 25:318-333, 16.

[4] J. Gilabert. "Enzyme Degradation and Physical and Chemical Characteristics of

Peach Bagte Pectin." Dissertation. Universitat de Lleida, 1998
[5]Álvarez, C. Ceballos, A. Caicedo, N.(2020). Pectin-to-Galacturonic Acid: Waste-to-
Bioproducts. Universidad icesi

[6] Molnár, E. Nemestóthy, N. & Bélafi-Bakó, K. (2010). Utilisation of bipolar

electrodialysis for recovery of galacturonic acid. Desalination. 1128-1131.

[7] Liu, S. (2020). Bioprocess Engineering Kinectics, Sustainability, and Reactor

Design (Tercera ed.). Nueva York, Estados Unidos: ELSEVIER.